One of the indications for BCR::ABL1 mutation testing in chronic myeloid leukemia (CML) is when tyrosine kinase inhibitor therapy (TKI) needs to be changed for unsatisfactory response. In this study, we evaluated a droplet digital PCR (ddPCR)-based multiplex strategy for the detection and quantitation of transcripts harbouring mutations conferring resistance to second-generation TKIs (2GTKIs). Parallel quantitation of e13a2, e14a2 and e1a2 BCR::ABL1 fusion transcripts enables to express results as percentage of mutation positive- over total BCR::ABL1 transcripts. We determined the limit of blank in 60 mutation-negative samples. Accuracy was demonstrated by further analysis of 48 samples already studied by next generation sequencing (NGS). Mutations could be called down to 0.5% and across 3-logs of BCR::ABL1 levels. Retrospective review of BCR::ABL1 NGS results in 513 consecutive CML patients with non-optimal response to first- or second-line TKI therapy suggested that a ddPCR-based approach targeted against 2GTKI-resistant mutations would score samples as mutation-negative in 22% of patients with warning response to imatinib but only in 6% of patients with warning response to 2GTKIs. We conclude ddPCR represents an attractive method for easy, accurate and rapid screening for 2GTKI-resistant mutations impacting on TKI selection, although ddPCR cannot identify compound mutations.

Droplet digital PCR for the detection of second-generation tyrosine kinase inhibitor-resistant BCR::ABL1 kinase domain mutations in chronic myeloid leukemia / Soverini S.; De Santis S.; Martelli M.; Monaldi C.; Castagnetti F.; Gugliotta G.; Papayannidis C.; Mancini M.; Bruno S.; Venturi C.; Machova Polakova K.; Ernst T.; Maar D.; Corner A.; Cavo M.. - In: LEUKEMIA. - ISSN 0887-6924. - STAMPA. - 36:9(2022), pp. 2250-2260. [10.1038/s41375-022-01660-8]

Droplet digital PCR for the detection of second-generation tyrosine kinase inhibitor-resistant BCR::ABL1 kinase domain mutations in chronic myeloid leukemia

Soverini S.;De Santis S.;Martelli M.;Monaldi C.;Castagnetti F.;Gugliotta G.;Papayannidis C.;Mancini M.;Bruno S.;Venturi C.;Cavo M.
2022

Abstract

One of the indications for BCR::ABL1 mutation testing in chronic myeloid leukemia (CML) is when tyrosine kinase inhibitor therapy (TKI) needs to be changed for unsatisfactory response. In this study, we evaluated a droplet digital PCR (ddPCR)-based multiplex strategy for the detection and quantitation of transcripts harbouring mutations conferring resistance to second-generation TKIs (2GTKIs). Parallel quantitation of e13a2, e14a2 and e1a2 BCR::ABL1 fusion transcripts enables to express results as percentage of mutation positive- over total BCR::ABL1 transcripts. We determined the limit of blank in 60 mutation-negative samples. Accuracy was demonstrated by further analysis of 48 samples already studied by next generation sequencing (NGS). Mutations could be called down to 0.5% and across 3-logs of BCR::ABL1 levels. Retrospective review of BCR::ABL1 NGS results in 513 consecutive CML patients with non-optimal response to first- or second-line TKI therapy suggested that a ddPCR-based approach targeted against 2GTKI-resistant mutations would score samples as mutation-negative in 22% of patients with warning response to imatinib but only in 6% of patients with warning response to 2GTKIs. We conclude ddPCR represents an attractive method for easy, accurate and rapid screening for 2GTKI-resistant mutations impacting on TKI selection, although ddPCR cannot identify compound mutations.
2022
Droplet digital PCR for the detection of second-generation tyrosine kinase inhibitor-resistant BCR::ABL1 kinase domain mutations in chronic myeloid leukemia / Soverini S.; De Santis S.; Martelli M.; Monaldi C.; Castagnetti F.; Gugliotta G.; Papayannidis C.; Mancini M.; Bruno S.; Venturi C.; Machova Polakova K.; Ernst T.; Maar D.; Corner A.; Cavo M.. - In: LEUKEMIA. - ISSN 0887-6924. - STAMPA. - 36:9(2022), pp. 2250-2260. [10.1038/s41375-022-01660-8]
Soverini S.; De Santis S.; Martelli M.; Monaldi C.; Castagnetti F.; Gugliotta G.; Papayannidis C.; Mancini M.; Bruno S.; Venturi C.; Machova Polakova K.; Ernst T.; Maar D.; Corner A.; Cavo M.
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11585/903554
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